The complementary roles of VAMP-2, -3, and -7 in platelet secretion and function.

Platelet secretion requires Soluble N-ethylmaleimide Sensitive Attachment Protein Receptors (SNAREs). Vesicle SNAREs/Vesicle-Associated Membrane Proteins (v-SNAREs/VAMPs) on granules and t-SNAREs in plasma membranes mediate granule release. Platelet VAMP heterogeneity has complicated the assessment of how/if each is used and affects hemostasis. To address the importance of VAMP-7 (V7), we analyzed mice with global deletions of V3 and V7 together or platelet-specific deletions of V2, V3, and global deletion of V7. We measured the kinetics of cargo release, and its effects on three injury models to define the context-specific roles of these VAMPs. Loss of V7 minimally affected dense and α granule release but did affect lysosomal release. V3-/-7-/- and V2Δ3Δ7-/- platelets showed partial defects in α and lysosomal release; dense granule secretion was unaffected. In vivo assays showed that loss of V2, V3, and V7 caused no bleeding or occlusive thrombosis. These data indicate a role for V7 in lysosome release that is partially compensated by V3. V7 and V3, together, contribute to α granule release, however none of these deletions affected hemostasis/thrombosis. Our results confirm the dominance of V8. When it is present, deletion of V2, V3, or V7 alone or in combination minimally affects platelet secretion and hemostasis.

[1]  A. Descoteaux,et al.  VAMP3 and VAMP8 Regulate the Development and Functionality of Parasitophorous Vacuoles Housing Leishmania amazonensis , 2022, Infection and immunity.

[2]  B. Storrie,et al.  Alterations in platelet secretion differentially affect thrombosis and hemostasis. , 2018, Blood advances.

[3]  T. Galli,et al.  Biomechanical Control of Lysosomal Secretion Via the VAMP7 Hub: A Tug-of-War between VARP and LRRK1 , 2018, iScience.

[4]  R. Flaumenhaft,et al.  The life cycle of platelet granules , 2018, F1000Research.

[5]  C. Moncman,et al.  Cellubrevin/vesicle-associated membrane protein-3-mediated endocytosis and trafficking regulate platelet functions. , 2017, Blood.

[6]  Smita Joshi,et al.  The nuts and bolts of the platelet release reaction , 2017, Platelets.

[7]  T. Galli,et al.  The SNARE VAMP7 Regulates Exocytic Trafficking of Interleukin-12 in Dendritic Cells , 2016, Cell reports.

[8]  T. Galli,et al.  VAMP-7 links granule exocytosis to actin reorganization during platelet activation. , 2015, Blood.

[9]  M. Yamakuchi,et al.  Abstract 181: VAMP8 Mediates Endothelial Granule Exocytosis , 2014, Arteriosclerosis, Thrombosis, and Vascular Biology.

[10]  A. Filipovich,et al.  Syntaxin-11, but not syntaxin-2 or syntaxin-4, is required for platelet secretion. , 2012, Blood.

[11]  C. Fader,et al.  ATP is released from autophagic vesicles to the extracellular space in a VAMP7-dependent manner , 2012, Autophagy.

[12]  Robert Flaumenhaft,et al.  Granule exocytosis is required for platelet spreading: differential sorting of α-granules expressing VAMP-7. , 2012, Blood.

[13]  C. Frassoni,et al.  TI‐VAMP/VAMP7 is the SNARE of secretory lysosomes contributing to ATP secretion from astrocytes , 2012, Biology of the cell.

[14]  Inés Rojo Pulido,et al.  VAMP3 is associated with endothelial weibel-palade bodies and participates in their Ca(2+)-dependent exocytosis. , 2011, Biochimica et biophysica acta.

[15]  Lydia Danglot,et al.  Multiple roles of the vesicular‐SNARE TI‐VAMP in post‐Golgi and endosomal trafficking , 2009, FEBS letters.

[16]  M. B. Mestre,et al.  TI-VAMP/VAMP7 and VAMP3/cellubrevin: two v-SNARE proteins involved in specific steps of the autophagy/multivesicular body pathways. , 2009, Biochimica et biophysica acta.

[17]  D. James,et al.  Variations in the requirement for v-SNAREs in GLUT4 trafficking in adipocytes , 2009, Journal of Cell Science.

[18]  Wade G. Regehr,et al.  Linking Genetically Defined Neurons to Behavior through a Broadly Applicable Silencing Allele , 2009, Neuron.

[19]  P. Roche,et al.  Mast cells possess distinct secretory granule subsets whose exocytosis is regulated by different SNARE isoforms , 2008, Proceedings of the National Academy of Sciences.

[20]  R. Tiedt,et al.  Pf4-Cre transgenic mice allow the generation of lineage-restricted gene knockouts for studying megakaryocyte and platelet function in vivo. , 2007, Blood.

[21]  W. Hong,et al.  Endobrevin/VAMP-8 is the primary v-SNARE for the platelet release reaction. , 2006, Molecular biology of the cell.

[22]  F. Mollinedo,et al.  Combinatorial SNARE Complexes Modulate the Secretion of Cytoplasmic Granules in Human Neutrophils1 , 2006, The Journal of Immunology.

[23]  T. Südhof,et al.  v‐SNAREs control exocytosis of vesicles from priming to fusion , 2005, The EMBO journal.

[24]  D. James,et al.  Combinatorial SNARE complexes with VAMP7 or VAMP8 define different late endocytic fusion events , 2004, EMBO reports.

[25]  A. Bernstein,et al.  Granule stores from cellubrevin/VAMP-3 null mouse platelets exhibit normal stimulus-induced release. , 2003, Blood.

[26]  T. Südhof,et al.  SNARE Function Analyzed in Synaptobrevin/VAMP Knockout Mice , 2001, Science.

[27]  T. Galli,et al.  Role of Tetanus Neurotoxin Insensitive Vesicle-Associated Membrane Protein (Ti-Vamp) in Vesicular Transport Mediating Neurite Outgrowth , 2000, The Journal of cell biology.

[28]  I. Martinez,et al.  Synaptotagmin VII Regulates Ca2+-Dependent Exocytosis of Lysosomes in Fibroblasts , 2000, The Journal of cell biology.

[29]  R. Scheller,et al.  Vamp-7 Mediates Vesicular Transport from Endosomes to Lysosomes , 1999, The Journal of cell biology.

[30]  W. Antonin,et al.  Mixed and Non-cognate SNARE Complexes , 1999, The Journal of Biological Chemistry.

[31]  S. Wong,et al.  Endobrevin, a novel synaptobrevin/VAMP-like protein preferentially associated with the early endosome. , 1998, Molecular biology of the cell.

[32]  T. Südhof,et al.  Cellubrevin is a ubiquitous tetanus-toxin substrate homologous to a putative synaptic vesicle fusion protein , 1993, Nature.

[33]  F. Benfenati,et al.  Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin , 1992, Nature.